ABNOH‐Linked Nucleotides and DNA for Bioconjugation and Cross‐linking with Tryptophan‐Containing Peptides and Proteins

• Published in: Chemistry : a European journal Vol. 30; no. 49; pp. e202402151 - n/a
• Main Authors: Spampinato, Ambra, Leone, Denise‐Liu', Pohl, Radek, Tarábek, Ján, Šoltysová, Markéta, Polák, Marek, Kádek, Alan, Sýkorová, Veronika, Řezáčová, Pavlína, Hocek, Michal
• Format: Journal Article
• Language: English
• Published: Germany Wiley Subscription Services, Inc 02-09-2024
• Subjects: Bioconjugations; Chemical synthesis; Conjugates; Cross-Linking Reagents - chemistry; Crosslinking; Deoxyribonucleic acid; DNA; DNA - chemistry; DNA biosynthesis; DNA polymerase; DNA probes; DNA-directed DNA polymerase; DNA-Directed DNA Polymerase - chemistry; DNA-Directed DNA Polymerase - metabolism; Enzymatic synthesis; N-oxyl radicals; Nucleotides; Nucleotides - chemistry; Oligonucleotides; Peptides; Peptides - chemistry; Proteins; Proteins - chemistry; Sodium nitrite; Substrates; Tryptophan; Tryptophan - chemistry; Nucleotides; Peptides; N-oxyl radicals; Oligonucleotides; Bioconjugations
• ISSN: 0947-6539; 1521-3765; 1521-3765
• DOI: 10.1002/chem.202402151

Reactive N‐hydroxy‐9‐azabicyclo[3.3.1]nonane (ABNOH) linked 2′‐deoxyuridine 5′‐O‐mono‐ and triphosphates were synthesized through a CuAAC reaction of ABNOH‐PEG4‐N3 with 5‐ethynyl‐dUMP or ‐dUTP. The modified triphosphate was used as substrate for enzymatic synthesis of modified DNA probes with KOD XL DNA polymerase. The keto‐ABNO radical reacted with tryptophan (Trp) and Trp‐containing peptides to form a stable tricyclic fused hexahydropyrrolo‐indole conjugates. Similarly modified ABNOH‐linked nucleotides reacted with Trp‐containing peptides to form a stable conjugate in the presence but surprisingly even in the absence of NaNO2 (presumably through activation by O2). The reactive ABNOH‐modified DNA probe was used for bioconjugations and crosslinking with Trp‐containing peptides or proteins.
Reactive probes based on ABNOH‐linked nucleotides, oligonucleotides or DNA were designed and synthesized for specific cross‐linking and bioconjugation with tryptophan‐containing peptides and proteins.